The Tissue Culture Laboratory

TCL Services




Custom Polyclonal Antibody Preparation
A single rabbit, which has been prescreened by ELISA for reactivity to the antigen, is injected five times. A total of four bleeds are taken and serum is analyzed twice for reactivity and specificity by ELISA. By request, additional rabbits can be used and additional bleeds can be performed.

Custom Hybridoma Preparation
Immunization
Five Balb/c mice are immunized four times and their antibody titers are examined by ELISA.

Cell Fusion
Myeloma cells are fused with spleenocytes from a single donor mouse.  The resulting cells are seeded onto ten 96 well cluster dishes and, after a period of growth, the conditioned supernatant media is collected for analysis.
 
 



 

Colony Selection
Each well of the fusion plates are analyzed by ELISA three times against the antigen.

Cloning
A single positive colony is cloned by limited dilution.  The clones are expanded and media is collected for analysis.

Expansion and Storage
Six clones are expanded to 1 X 10⁷ cells and stored in liquid nitrogen at 3 x 10⁶ cells, 3 vials per clone.
 

Antibody / Protein purification by Fluid Phase Liquid Chromatography (FPLC)
Information available upon request.
 

ELISA
Information Available Upon Request
 

Monoclonal Antibody Production
In vitro production of antibodies from existing hybridomas

A continuous cell culture system is available for the efficient expression of antibodies in vitro. A ten to fifty-fold increase in antibody concentration is exhibited over traditional cell culture methods. Additional information is available upon request.
 
Cell Line Maintenance
Information available up request
 

Cell Storage in Liquid Nitrogen
A single cell line is expanded to 2 x 10⁷ cells and stored in liquid nitrogen at 3 x 10⁶ cells/ml x 6 vials.


Establishment of Primary Cultures
Charges are based on the reagents and complexity of the particular protocol.
 

Mycoplasma Screening
The sample cell lines are expanded for one week in antibiotic free medium.  Then, ICN Biomedicals' Immuno-Mark Myo-Test Kit^TM is used to screen for mycoplasma.


Mycoplasma Contamination Treatment
A contaminated cell line is treated with antibiotics specific for mycoplasma. More information is available upon request.
 

Mailing Address:
The Department of Pharmacology and Toxicology
School of Pharmacy
University of Kansas
1251 Wescoe Hall Drive
5064 Malott Hall
Lawrence, KS 66045-7582